A Study Of The Effects Of MIFsiRNA On The Biological Characters Of Colorectal Cancer Cellines

Background:Multiple functions have been ascribed to macrophage migration inhibitory factor(MIF),such as playing a pivotal role in signal regulation,inflammatory,host innate andacquired immunity.MIF also controls cell proliferation and differentiation,migration andpromotes tumor neovascularization,progression and metastasis.The small interfering RNA of macrophage migration inhibitory factor(MIFsiRNA) isa matter which is a chemical synthesis to aim directly at MIF ,combining with th-emRNA of MIF specifically,leading to the sequence of mRNA of MIF be resolved,then the proteinum of MIF'translation be depressed ,and the biological function of MIF be inhibited.therefore,we supposed that,in colorectal cancer cell lines, the proliferation and invasion be restrained to ,and the apoptosis be increased to by MIFsiRNA.Objective:To study the effects of MIFsiRNA.A matter of chemical synthesis,on the biologicalcharacter of colorec tal cancer cell lines,and explore the possible mechanisms,then we canopen up new channels for the prevention and treatment for the metastasis of colorectaltumor.Methods:1.The fluorimetric experimental group was transfected by the FAMsiRNA with the concentration of 100 nM/ml,while the blank group is not add to any siRNA.2.In the colorectal cancer cell lines ,the positive control group was transfected by theMIFsiRNA with the range of concentration from the 10 nM/ml to 100 nM/ml,the negativecontrol group was treated by nonspecific siRNA with the same concentration as positivecontrol group,and the blank control group had no any siRNA be added to .3.MIF,CD74,Caspases3,Caspases8,E-Cadherin,Tiam1 were detected in mRNA lineswith RT-PCR after the colorectal tumor cells of CT-26 were transfected in order withMIFsiRNA,nonspecific siRNA,and the blank control group had no any siRNA be addedto .4.MTT was used to detect the effect on the proliferation of colorectal cancer cell lines ofCT-26 between MIFsiRNA group,nonspecific siRNA group and blank control group.5.When CT-26 cells were transfected by MIFsiRNA and nonspecific siRNA,the expressionof proteinum of MIF in culture solution was deteced by ELISA6.The effect on invasion,in vitro of CT-26 cells, been detected by transwell betweenMIFsiRNA and nonspecific siRNA7.Flow cytometer was used to evaluate the effect of the apoptosis of CT-26 cells,whencolorectal cancer cells were transfected by MIFsiRNA and nonspecific siRNA.8.Western blot was used to evaluated the expression of proteinum in vivo of MIF,CD74,Caspases8.ResultResult:1,The fluorimetric siRNA was transfected in vivo.2,MIFsiRNA inhibited the proliferation of CT-26 colorectal cells,which depended ontime-dose,but the nonspecific siRNA and the blank control group had no effect on CT-26cells.3,MIFsiRNA with the concentration of 100 nM/ml transfected colorectal cancer cellsleading the expression of MIF,CD74,Tianm1 to decrease ,and the E-Cadherin,Caspases3,Caspases8 to increase,that had significant deviation comparing with thenonspecific siRNA group and blank control group,whereas ,the nonspecific siRNA groupand blank control group had no difference. 4,In vitro ,the proteinum of MIF were depressed after 100 nM/ml MIFsiRNA treatment for24 hours,and the blank group and the nonspecific siRNA had no influence with theexpression of the MIFs'proteinum.5,The colorectal cancer cells penetrated polycarbonate membrane notable reduction after100 nM/ml MIFsiRNA treatment for 24 hours,and ,in comparation with the blank groupand the nonspecific siRNA group,it had significant difference, the nonspecific siRNA groupand blank control group had no difference.6,In comparation with the blank group and the nonspecific siRNA group ,the apoptosisincreased after 100 nM/ml MIFsiRNA treatment for 24 hours,and the nonspecific siRNAgroup and blank control group had no difference.7,In vivo,the expression of the proteinum of MIF,CD74 were decreased,and the Caspases8was increased, after 100 nM/ml MIFsiRNA treatment for 48 hours,that comparing withnonspecific siRNA and blank group had significant deviation,but the nonspecific siRNAand the blank group had no discrepancy.ConclConclusions:1,MIFsiRNA depressed the proliferation of colorectal cancer cells of CT-26.The possiblemechanism may be MIFsiRNA degrade the combination between MIF and CD74,thendown regulate the MEK/ERK path,and the cells'proliferation are down-regulated.2,MIFsiRNA depressed the invasion of colorectal cancer cells of CT-26.The possiblemechanism may be MIFsiRNA degrade the expression of MIF,then up regulate theE-Cadherin's expression,and down regulate the Tiam1's expression.3,MIFsiRNA depressed the invasion of colorectal cancer cells of CT-26.The possiblemechanism may be MIFsiRNA degrade the expression of MIF,then up regulated theexpression of Caspases3 and Caspases8.